ABSTRACT
In this study, the rapid resolution liquid chromatography-quadrupole time-of-flight mass spectrometry (RRLC-QTOF/MS) was used to profile the metabolites of urine samples from chronic heart failure (CHF) patients and healthy controls to find the differential metabolites which could provide the scientific evidence to explain the pathogenesis of the disease and supply a better therapy plan.Urine samples from 15 CHF patients (age (62.27±3.14) years) and 15 healthy controls (age (65.41±4.63) years) were analyzed by RRLC-QTOF/MS.After processing the data, the multivariate statistical analysis (principal component analysis, PCA) was performed to find the potential biomarkers.Result showed that urine samples of CHF patients were successfully distinguished from those of healthy controls.Two significantly differentially expressed metabolites, uridine and alanyltryptophan, were found and identified as potential biomarkers.The result showed that the LC-MS based metabolomics approach had good performance to identify potential biomarkers, and the disorder of uracil metabolism and Tryptophan metabolism may play an important role in the mechanism of CHF.
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A rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometric (RRLC-Q-TOF-MS) method was established and optimized for the analysis of pharmacokinetic behavior of ginsenoside Rb2 in rats by intravenous injection administration.The metabolism of ginsenosides Rb2 in vivo rat was also explored.In the experiment,Agilent SB C18 column was selected for the sample separation with 0.1％ aqueous formic acid solution as mobile phase (A) and acetonitrile as mobile phase (B) at a flow rate of 0.2 mL/min,and the injection volume was set to 5 μL.Q-TOF-MS was carried out in electron pray ionization (ESI) negative ion mode.The limit of quantification (LOQ,S/N =10) and limit of detection (LOD,S/N=3) were 0.10 and 0.08 μg/mL,respectively,and the linear range was 0.1-1.26 μg/mL.The experiment results showed that the concentration-time profile of ginsenoside Rb2 conformed to a two-compartment pharmacokinetic model after intravenous administration for rats.The mean plasma elimination half-lives were (23.58±1.10) min (t1/2α),(1306.55±147.23) min (t1/2β) for Rb2.By analyzing the urine of rats after intravenous administration and the fecal samples after oral administration of ginsenoside Rb2,it was found that the metabolites were M6,M2 (CY),F2,and C-K.
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A rapid resolution liquid chromatography quadruple time-of-flight mass spectrometric ( RRLC-Q-TOF/MS) method combined with multivariate statistical analysis was applied to investigate the changes of endogenous metabolites in murine urine before and after ultraviolet B ( UVB ) irradiation for the purpose of discussing the physiological mechanism of acute injury caused by UVB radiation. A narrow-band UVB ( NB-UVB) (TL-01, peak value 312nm) was used to establish the acute light damage model. The urine samples were centrifuged before four times dilution treatment, subsequently the diluted urine samples were separated on a Supelco Ascentis Express C18 column using water (0. 1% formic acid) and acetonitrile as mobile phase by gradient elution. The differences metabolites with major contribution for grouping were found out based on the metabolic profiling analysis of principal component analysis ( PCA) and cluster analysis ( CA) , which could illustrate their possible mechanism of actions by means of relevant pathways. A prediction model was built to investigate the forecasting ability of the acute photo damage induced by UVB irradiation through the partial least square discriminant analysis ( PLS-DA ) . The results of multivariate statistical analysis showed that the blank control group was separated from UVB model group quite well, 11 endogenous metabolites were identified as the potential biomarkers through comparison with the database, tandem mass spectrum data and standard substance, which indicated the UVB radiation may affect the sphingolipid metabolism, nucleic acid metabolism, linoleic acid metabolism and amino acid metabolism pathways. These different metabolites could be helpful for diagnosing the light damage induced by UVB radiation
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In this study, the rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry ( RRLC-QTOF/MS ) was used to profile the metabolites of urine samples from Childhood Pneumonia ( CP) patients and healthy controls and find the potential biomarkers which can support evidence to early diagnose and cure the disease. Choose 10 CP patients ( age 47. 72 ± 2. 35 months) and 10 healthy controls ( age 46 . 65 ± 1 . 97 months ) . The urine samples were analyzed by RRLC-QTOF/MS and then the resulting data matrices were analyzed by principal components analysis ( PCA ) to find the potential biomarkers. Urine samples of CP patients were successfully distinguished from those of healthy controls. A total of two significantly changed metabolites have been found and identified as potential biomarkers. It is suggested that the disorder of purine metabolism and amino acid metabolism may play an important role in the mechanism of CP.
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A rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometric ( RRLC-QTOF/MS) method was used to profile the metabolites of urine samples from atherosclerosis ( AS) patients and healthy controls and find the differential metabolites which could provide the scientific evidence to explain the pathogenesis and early disease diagnose. In the study, 15 AS patients ( age46. 84±2. 41 years) and 15 healthy controls ( age45 . 72±1 . 93 years ) was screened out by VaSera VS-1000 . The urine samples were analyzed by RRLC-QTOF/MS and the resulting data matrices were analyzed by multivariate statistical analysis ( Principal Component Analysis, PCA ) to find the potential biomarkers. The results showed that the urine samples of AS patients were successfully distinguished from those of healthy controls. Besides, a total of two significantly changed metabolites, uric acid and Guanidineacetic acid, had been found and identified as potential biomarkers, which suggested that the disorder of purine metabolism and amino acid metabolism played an important role in the mechanism of AS.
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AIM@#To analyze the composition of the Chinese herbal medicine Sanjie Zhentong Capsule (SJZTC) and test the therapeutic efficacy of each component in a rat model of endometriosis.@*METHODS@#A rapid resolution liquid chromatography (RRLC) method coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS) has been developed for the analysis of SJZTC. Two main ingredients, Drac(h)onis sanguis and saponin, were tested in the endometriosis model. Sixty Lewis female rats were in the estrous cycle stage when endometriosis was experimentally initiated by peritoneal implantation of endometrial tissue. Four weeks later, a second laparotomy was performed and implant volumes were measured. After that, the implanted rats were randomized into five study groups: control group (treatment with saline), anastrozole group (treatment with anastrole, 18 μg per day), loureirin A group (treated with loureirin A, 97.2 mg), ginsenoside Re group (treated with ginsenoside Re, 64.8 mg), and SJZTC groups (treated with SJZTC, 86.4 mg) administered once a day for 4 weeks via gastric gavage. After four weeks of treatment, a third laparotomy was performed, implant volumes were re-measured, and the levels of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) were tested.@*RESULTS@#A total of 38 compounds including, both the target and unknown compounds, were rapidly predicted in the capsule extract by the developed method. Compared with the control group, the anastrozole group, loureirin A group, ginsenoside Re group, and SJZTC treated group showed smaller implant volumes, as well as lower levels of VEGF and TNF-α in the peritoneal focus (P < 0.01 for all comparisons). Furthermore, parameters in the groups treated with SJZTC, loureirin A and ginsenoside Re were significantly better than the control group and the anastrozole group. These results indicate that SJZTC and its two main components are effective in reducing the development of endometriosis.
Subject(s)
Animals , Female , Humans , Rats , Capsules , Chemistry , Disease Models, Animal , Drugs, Chinese Herbal , Chemistry , Endometriosis , Drug Therapy , Rats, Inbred LewABSTRACT
Objective: A novel method for the analysis of alkaloids in Nelumbo nucifera leaves was established by SPE-RRLC-Q-TOF mass spectrometry. Methods: The crude sample was extracted by 1% HCl with ultrasound-assisted extraction, then purified by SPE column, and eluted with ammonia-methanol (5:95). After concentration, the residue was dissolved by methanol solution. The real sample was analyzed by RRLC-Q-TOF. A Welch Materials C18 column was applied in the RRLC separation using acetonitrile and water as mobile phase. The elutes were detected by Q-TOF to obtain the MS spectra with extract molecular weights under positive ion mode. Results: Nine alkaloids were identified. Conclusion: This method can be used to rapidly determine the alkaloids of N. nucifera leaves.
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Objective: To analyze the constituents and metabolites of Radix Sa poshnikoviae (RS) in rat plasma and urine by rapid-resolution liquid chromatography-time of flight mass spectrometry (RRLC-TOF/MS), so as to explore the active ingredients and metabolites of RS in vivo. Methods: The separation was performed on a Angilent Zorbax Extend-C14 (5 μm, 250 mm x 4.6 mm id) column, with a methanol-water mobile phase system used for gradient elution. Time-of-flight mass spectrometer (TOF/MS) was applied for qualitative analysis under positive ion mode. Based on the accurate molecular weight of TOF/MS detection and the compound list of RS established previously, the constituents and metabolites of RS in different matrix in vivo were identified. Results: Six constituents of RS were identified in the plasma, sucrose, prim-O-glucosylcimifugin, cimifugin, nodakenetin, 5-O-methylvisamminol, and 3′-O-i-butyrylhammaudol. Eight constituents were identified in the urine, prim-O-glucosylcimifugin, divaricatacid, cimifugin, 4′-O-glucosyl-5-O- methylvisamminol, (3S)-2,2-dimethyl-3, 5-dihydroxy-8-hydroxymethyl-3, 4-dihydro-2H, 6H-benzo-[1, 2-b: 5, 4-b′] dipyran-6-one, 5-O-methylvisamminol, see-O-β-D-glucosylhammaudol, and wogonin. Two metabolites were identified in the urine, glucuronide of cimifujin and an isomer of it. Conclusion: The present method is reliable and effective for identifying compounds of RS in vivo, and it can provide a reference and evidence for the further pharmacodynamics experiments.
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A method was developed for the determination of zeranols (α,β-zearalanol,α,β-zearalenol,zear-alanone,zearalenone) with RRLC-MS/MS in the plant tissue. The samples were extracted with acetonitrile and reextracted with aqueous alkaline solution,cleaned up with MAX column and then determined by RRLC-MS/MS using multiple reaction monitoring ( MRM) scan mode. The results showed that the working curves were linear in the range of 0 -20 μg/kg. The limits of detection ( LOD) of zeranols were 0.5 μg/kg and the limits of quantification (LOQ) was 1. 0μg/kg. Extraction recoveries for zeranols ranged from 75. 8% to 105.4% and RSD was between 2.4% and 12.1%. The method is suitable for the determination of zeranols in the plant tissue.